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7;there were 32 b ands in tumor group. ③SDS PAGE showed that the number of protein bands with relative molecular mass of 77 000,50 000-52 000,38 000-30 000 increased in the tumour group. Conclusion: In the saliva of periodontitis indivauals there are more basic proteins,the relative molecular mass of the prot ein in the saliva of patients with tumor is different from that of health contro ls.
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objective:To study the growth inhibition effect of 5azaC on human salivary gland cell line HSG. Methods:HSG cells were exposed to 5-azaC at 5?10~ -6 mol/L and 10?10~ -6 mol/L respectively for 3 days. The proliferation of in vitro cultured HSG cells was studied by cell counting. The in vivo growth of HSG cells was investigated by tumor weight measurement in nude mouse models of HSG tumor induced by transplantation of the cells subcutaneously.Results:5 azaC inhibited HSG cell proliferation by 85% and 95% respectively at above mentioned doses. In the 3-week tumor growth study, the growth of the tumor induced by 5?10~ -6 mol/L 5azaC treated cell was inhibited by 74.8%.Conclusion:5azaC can inhibit the growth of HSG cells in vitro and in vivo.
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Objective:To estimate the effect of HSV1-TK/GCV suicide gene therapy on human salivary gland epidermoid carcinoma cell line MEC-1. Methods: Expression vector G 1NAtK containing HSV-TK cDNA was transfected into MEC-1 cells.After transfection, the cells were selected by G418 for two weeks. The integrated gene and mRNA were detected with PCR and RT-PCR. The cytotoxicity and bystander effect were estimated by MTT and typan blue exclusion assay. The morphological changes after GCV treatment were observed with HE and 33258 stain and in situ cell apoptosis detection kit. Results: The 404 bp DNA fragment was amplified through PCR and RT-PCR in the transfected cells respectively. TK positive clone was named MEC-1/TK. The sensitivity of MEC-1/TK to GCV was 1 000 times more than that of parent MEC-1 cells.More than 90% of MEC-1 cells were killed by 10 ?g/ml of GCV when only 10% of MEC-/TK cells were present. The morphological changes included shrinking,detaching and floating of the cells. Some of the cells showed nucleus condensation and breakage of nucleus. A lot of cells showed nucleus positive in in situ apoptosis detection. Conclusion: HSV1-TK/GCV can confer MEC-1/TK cell killing efficiently. MEC-1/TK also has strong bystander effects. HSV1-TK/GCV system confers its effect, in part, by inducing apoptosis in TK positive cells.
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Objective: To investigate the induction of Tca8113 cells to apoptosis by fadd gene.Methods: RT PCR and recombinant PCR were used to amplify human fadd gene and cloned into expression vector pcDNA3 and pIRES2 EGFP, then transfered into Tca8113 cells. The growth and apoptosis of the cells were tested by cell counting,fluorescent microscopy, electron microscopy and flow cytometery. Results: fadd gene was obtained and transfered into Tca8113 cells. After transfection of the gene the growth of the cells was inhibited by 25%～52%, cell number in G 1 phase increased and that in S phase decreased. Apoptosis of the cells was observed. Conclusion: fadd gene can effectively inhibite cell growth and induce Tca8113 cells to apoptosis.
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Objeact:To evaluate the effects of the exogenous PTEN gene on morphology of the highly metastatic mucoepidermoid carcinoma cell line M3SP2. Methods: Morphological observation of vehicle transfected M3SP2-pBp cells and PTEN transfected M3SP2-PTEN cells was performed with inverted microscope, HE staining and optical microscope, scanning electronic microscope and transmisson electronic microscope. Results: Compared with the control cells of M3SP2-pBp, the exogenous PTEN expressing cells M3SP2-PTEN showed morphological changes, such as vacuole denaturalization, shrinkage, less microvillus, chondriosome swelling, lysosome amalgamation, and chromatin agglutination. Conclusion: The exogenous PTEN gene may induce denaturalization, necrosis, and apoptosis of the highly metastatic mucoepidermoid carcinoma cells.
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Objctive: To study the inhibitory effects of homoharringtonine(HHT) or methoxsalen (8-MOP) or in combinnation on humen highly metastatic mucoepidermoid carcinoma Mc3 cells. Methods: Mc3 cells were exposed to the drugs or their combination at various concentrations. The inhibitory effects were tested with MTT assay. Results: The IC 50 values of HHT and 8-MOP were 79.43 ng/ml and 5 980 ng/ml respectively, HHT at the doses of 1-320 ng/ml combined with 8-MOP at 800, 4 000 and 20 000 ng/ml the CI 50 values were 0.76?0.33 and 0.15 respectively.Conclution: The data indicate that combination of HHT with 8-MOP is synergic in the inhibition of Mc3 cell growth.
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Objective To evaluate the biocompatibility of ZSM-5 zeolite and provide an experimental basis for developing the first-aid hemostatic agent.Methods According to Chinese evaluation standards on medical devices and biological tests,the cytotoxicity in vitro,hemolysis test,acute toxicity of system,pyrogen test,intracutaneous stimulation,sensitization and micronucleus test were studied in ZSM-5 zeolite.In order to find out the side-effect of the zeolite granules' remains left in the wounds to body,muscle implantation test was studied.Results There were no obvious cytotoxity,hemolysis reaction.Acute tocicity,pyrogen reaction,intraeutaneous stimulation,sensitization and potential mutagenesis in themicronucleus test were observed.Their results were a11 consistent with the Chinese biological evaluation of medical devices.Obvious inflammatory reaction was observed when ZSM-5 zeolite was implanted in muscle for 12 weeks.Conclusion The ZSM-5 zeo1ite has reliable biocompatibility.But zeolite can cause inflammatory reaction when it is remained in the wound surface for long term.